Compositions and methods for making and using three-dimensional tissue systems

Abstract

The present disclosure provides methods, compositions, and devices for making and using three-dimensional biological tissues that accurately mimic native physiology, architecture, and other properties of native tissues for use in, among other applications, drug testing, tissue repair and/or treatment, and regenerative medicine.

The invention claimed is: 
     
       1. A bioreactor comprising:
 a device having a well configured for growing a tissue from cells seeded therein, wherein the well has a bottom; and 
 at least two elastic sensing elements disposed across the well such that there is a gap between the sensing elements and the bottom of the well, wherein the sensing elements are configured to: (a) permit attachment of the tissue formed therebetween, thereby suspending the tissue above the bottom of the well, and (b) deform in response to the contractile force exerted on the sensing elements by the tissue, thereby simulating a physiological environment that is native to the tissue and/or permitting measurement of the contractile force. 
 
     
     
       2. The bioreactor of  claim 1 , comprising 2 to 25 sensing elements per well. 
     
     
       3. The bioreactor of  claim 1 , wherein the bioreactor is a multi-well plate comprising 6 wells, 12 wells, 24 wells, 96 wells, 384 wells, or 1536 wells. 
     
     
       4. The bioreactor of  claim 1 , wherein the sensing elements comprise a natural materials selected from the group consisting of collagen and collagen derivatives, animal intestines, cellulose and cellulose derivatives, proteoglycans, heparin sulfate, chondroitin sulfate, keratin sulfates, hyaluronic acid, elastin, fibronectin, laminin, fibrin, chitosan, alginate, matrigel, and combinations thereof. 
     
     
       5. The bioreactor of  claim 1 , wherein the sensing elements comprise a polymer selected from the group consisting of synthetic and biologic, and wherein the polymer is degradable or nondegradable. 
     
     
       6. The bioreactor of  claim 5 , wherein the polymer is at least one of polylactic acid, poly(lactic-co-glycolic) acid, poly(caprolactone), polyglycolide, polylactide, polyhydroxobutyrate, polyhydroxyalcanoic acid, chitosan, hyaluronic acid, a hydrogels, poly(2-hydroxyethyl-methacrylate), poly(ethylene glycol), poly(L-lactide) (PLA), poly(dimethysiloxane) (PDMS), poly(methylmethacrylate) (PMMA), poly(glycerol sebacate), poly(octamethylene maleate (anhydride) citrate) (POMaC), POMaC without citric acid, poly(ε-caprolactone), polyurethane, silk, a nanofabricated material, a co-polymer, a blended polymer, or a combination thereof. 
     
     
       7. The bioreactor of  claim 6 , wherein the polymer is POMaC. 
     
     
       8. The bioreactor of  claim 1 , wherein the sensing elements comprise a polymer which is doped with a nanostructure. 
     
     
       9. The bioreactor of  claim 1 , wherein the sensing elements comprise at least one of an intestinal material, monocryl, polyglycolide, prolene, polyglactin, polydioxanone, polypropylene, nylon, polyester, or a combination thereof. 
     
     
       10. The bioreactor of  claim 1 , further comprising a cell seeded in the well. 
     
     
       11. The bioreactor of  claim 10 , wherein the cell seeded in the well is at least one of a cardiomyocyte, a skeletal muscle cell, a hepatocyte, a renal cell, a chondrocyte, a skin cell, a contractile cell, a blood cell, an immune system cell, a germ cell, a neural cell, an epithelial cell, a hormone secreting cell, a bone marrow cell, a stem cell, a tumor cell, a smooth muscle cell, an endothelial cell, a fibroblast, an adipose derived stem cell, a mesenchymal stem cell, a progenitor cell, or combinations thereof. 
     
     
       12. The bioreactor of  claim 10 , wherein the cell seeded in the well is a cardiomyocyte. 
     
     
       13. The bioreactor of  claim 1 , further comprising a tissue formed therein. 
     
     
       14. The bioreactor of  claim 1 , wherein the well is configured to have a longitudinal axis. 
     
     
       15. The bioreactor of  claim 14 , wherein the sensing elements have an orientation that is perpendicular, parallel, or diagonal relative to the longitudinal axis of the well. 
     
     
       16. The bioreactor of  claim 1 , further comprising electrode configured to create an electrical current across the well of the bioreactor. 
     
     
       17. The bioreactor of  claim 1 , wherein the sensing elements comprise a polymer whose mechanical properties are tunable during a polymerization reaction. 
     
     
       18. The bioreactor of  claim 1 , wherein the sensing elements are porous, thereby permitting delivery of nutrients and growth factors to the tissue. 
     
     
       19. The bioreactor of  claim 1 , wherein the sensing elements have an elasticity from about 20 kPa to 0.5 MPa. 
     
     
       20. The bioreactor of  claim 1 , wherein the sensing elements are polymer wires. 
     
     
       21. The bioreactor of  claim 1 , wherein the cells are seeded in a hydrogel that comprises polyvinyl alcohol, sodium polyacrylate, an acrylate polymer, agarose, methylcellulose, or hyaluronan. 
     
     
       22. A method for measuring the effect of a test agent on the contractile force of a tissue using the bioreactor of  claim 1 , comprising:
 (a) measuring a first contractile force of the tissue in the bioreactor of  claim 1  before exposure to the test agent; 
 (b) contacting the tissue with the test agent under conditions sufficient for the test agent to affect the contractile force; 
 (c) measuring a second contractile force of the tissue after exposure to the test agent; 
 (d) determining whether the test agent affects the contractile force by comparing the first contractile force of (a) with the second contractile force of (c), 
 wherein measuring the first or second contractile force comprises measuring the amount of movement imposed by the tissue on the sensing elements from a first position to a second position. 
 
     
     
       23. The method of  claim 22 , wherein the test agent is selected from the group consisting of a small molecule, an antibody, an ion, a protein, a peptide, a lipid, DNA, RNA, a virus, bacteria, a microparticle, a nanoparticle, a therapeutic agent, and a toxin. 
     
     
       24. A method for evaluating the safety of a test agent on a tissue, comprising:
 (a) contacting the tissue of the bioreactor of  claim 1  with the test agent; 
 (b) measuring the effect on one or more physiological parameters indicative of safety; 
 (c) comparing the physiological parameters in (b) to the same physiological parameters measured from a control bioreactor not exposed to the test agent,
 wherein a statistically significant change in the physiological parameters in (b) as compared to the same physiological parameters measured from the control bioreactor indicates that the test agent lacks safety. 
 
 
     
     
       25. A method for fabricating the bioreactor of  claim 1  for cultivation of a tissue, comprising the steps of:
 (a) microfabricating a surface comprising a well configured for growing a tissue from cells seeded therein; and 
 (b) affixing at least two sensing elements to each well, wherein the sensing elements are configured to become encapsulated by the tissue once formed in the well. 
 
     
     
       26. A method comprising:
 (a) growing an implantable tissue in the bioreactor of  claim 1 ; and 
 (b) providing the implantable tissue for use in treating a damaged and/or diseased tissue.